Estrogenic substances



Patented May 7, 1946 UNITED STATES PATENT OFFICE ESTROGENIC SUBSTANCES Ralph C. Tallman. Westileld, N. 1., and Alfred 1!.

Stuart, New York, N. Y., 8: Co., New York, N. Y., a corporation of New York No Drawing. Application May 30, 1944, Serial No. 588,089

4 Claims. (Cl- 300-810) licular phase of the menstrual cycle in human females. Ordinarily such substances are secreted by the ovaries and the physiological changes brought about by the normal functioning of these ovarian secretions in conluction with other bodily functions.

However, due to disease, illness, surgical intervention, or many other causes, the normal functioning of the ovaries, and the consequent secretion of estrogem'c substances, may be absent or impaired. This condition may, in turn, give rise to other types of physical and mental illnesses. In such instances it often becomes advisable or necessary to administer therapeutically-for example, by injection or by feedingestrogenic substances produced outside the body of the patient to substitute for the absent or diminished natural secretions.

For industrial elaboration, estrogenic substances may be obtained from natural sources, such as the excretions of pregnant female mammals. FOr example, the urine of pregnant female horses may be collected and subjected to various processes designed to concentrate and purify the estrogenic substances contained therein, and thus to isolate them in a condition suitable for therapeutic administration in the manners and for the purposes named above. Such procedures are devious, time-consuming, and expensive.

Th s invention describes artificial estrogenic substances which may be prepared from simple chemical compounds. When the products described in this invention are employed, no recourse to estrogenic substances from natural sources is necessary. The compounds described below will induce physiological changes like those brought about by estrogenic substances from natural sources as outlined above. These compounds may be produced simply and inexpensively in commercial quantities, and may be used therapeutically for treatment of conditions arising from absence or diminution of the bodily produced estrogenic substances.

This invention is based on the discovery that estrogenic and bactericidal properties are possessed by compounds having the structure .xo 0 a a a ox in which each X is H, alkyl, acyl of aliphatic or aromatic type, or an inorganic ester radical such as 803K or POsHz which may be obtained by condensing one of these compounds having 0H terminal groups with an inorganic acid; and R is H or alkyl, at least one B. being alkyl; in which the OK groups may be symmetrical or not, and may be the same or diflerent; and in which each alkyl It may be the same or different.

The invention is further based on the further discovery that compounds having the following type formulae are especially and uniquely beneficial, and it is a further object of the invention to utilize said compounds as estrogenic and bactericidal compounds.

TYPE I H n a ox in which X is H, alkyl, or acyl, and R is alkyl.

TYPI II x0 I: a n ox 0 0 0 1'1 in which X is H, alkyl, or acyl, and R. is alkyl.

HRR

x0 R H R ox 0 0 0 0 y in which X is H, alkyl, or acyl, and each R is alkyl.

TYPI V in which x is H, alkyl, or acyi, and each a is alkyl.

The OX radical may be in the ortho, meta, or para position in each of the foregoing types, and need not be symmetrically positioned.

Examples of the production of compounds of the above types are as follows:

Example 1.-Production of Type I Specific members of the series illustrated by pe I may be prepared in the following manner: 43 grams of p-methoxyacetophenone and 45 grams of anisaldehyde are dissolved in 150 cc. of alcohol. The solution is stirred vigorously, and 150 cc. of 10% aqueous sodium hydroxide solution is gradually added in small portions. The product separates slowly from the solution and becomes crystalline on the addition of seed. stirring is continued for two hours after the addition of sodium hydroxide is complete, and the mixture is then allowed to stand 12 to 24 hours at C. The p,p'-dimethoxy chalcone thus produced (see Step I below) is removed by filtration, washed with ice-cold alcohol and water, and may be purified by crystallization from about 400 cc. of alcohol, from which it separates in bright yellow crystals M. P. 100-101 C.

The D.P'-dimethoxy chalcone is reacted (see Step II below) with ethyl magnesium bromide as follows: A solution of 31 grams of ethyl magnesium bromide in 450 cc. dry ether is maintained at 10 C. and vigorously stirred while 20 grams 'of the chalcone is added in small quantities. The cooling bath is then removed, and stirring continued for two and one-half hours at room temperature. The reaction mixture is poured into 1200 cc. ice water containing 150 cc. hydrochloric acid, after which the ether layer is separated and the aqueous layer extracted once with ether. The ether is evaporated from the combined solutions and the product which remains is purified by distillation under high vacuum and crystallization from alcohol. The white crystals of the product, 1,3-di (p-methoxyphenyl) -pentanone-1 (where R is the ethyl group) have a melting point of 70 C. This compound may be converted into 1,3-di (p-methoxyphenyl) pentane as follows:

9 grams of the pentanone are mixed with 60 grams of amalgamated zinc, 45 cc. water, 120 cc. hydrochloric acid, 35 cc. of toluene, and 2 cc. acetic acid, and the reaction mixture heated under refiux at the boiling point for twenty-eight hours (see Step III below). It is then cooled and extracted with ether, the ether layer separated, and all solvents removed therefrom by evaporation If reduction is not complete, as shown by separation of crystals or by failure of the residue to boil at a constant temperature, the same process may be repeated until reduction is complete. The final product of this step, 1,3-di (p-methoxyphenyl) -pentane may be purified by distillation, and is a colorless liquid which boils at 160-165 C. under 1 mm. pressure. It may be converted into the corresponding phenolic product as follows:

2.0 grams of the 1,3-di (p-methoxyphenyl) pentane are hydrolyzed by dissolving in 6 cc. glacial acetic acid, to which solution 15 cc. of 57% aqueous hydriodic acid is added. The reaction mixture is maintained under reflux at the boiling point for 15 minutes, then 4 cc. of glacial acetic acid and 5 cc, of the hydriodic acid solution are added, and refluxing continued for a total of four hours. The mixture is poured into water, the solution made alkaline with excess potassium hydroxide, and filtered with the aid of charcoal. The phenolic product thus obtained (see Step IV below) is precipitated by acidification of the filtrate, extracted with ether, and, after the evaporation of the ether, distilled slowly at C. under high vacuum. The distillate is a colorless glass-like product which solidifies on long standing. This compound is 1,3-di-(p-hydroxyphenyl) -pentane, where R is the ethyl radical.

PRODUCTION or TYPE I Step I onloOfi-cm oHlo-Ogw CHaO-OE-CIECHOOCH:

Step II cmo-Og-cmcn-Qmom EtMgBr omo-O- -omoH-Oocm 0 Et Step III CH O- -CCHaCH- OCH; (reduction) omo-Oom-om-on-O-oom StepIV CmoO-CHrCHrLJiI-OO on, -v (hydrolysis) no-oom-cnz-c lnoon Example 2.-Pr0duction of Type II Specific members of the series illustrated by Type 11 may be prepared in the following manner: 16 grams of anisaldehyde and 22 gram of p-methoxybutyrophenone are thoroughly mixed, and a stream of dry hydrogen chloride gas bubbled into the mixture until the total weight is increased by 3.5 grams. The reaction mixture, which rapidly becomes very dark red, is shaken until it becomes pasty, due to the separation of the product, and is then allowed to stand for filteen hours. The entire mixture is dissolved in benzene and the benzene solution extracted with water, with sodium carbonate solution and again with water. The benzene is then evaporated and the residue maintained at 150 C. until evolution of hydrogen chloride has ceased, after which it is subjected to distillation under reduced pressure. After a small amount of lower boiling forerun, which is discarded, the desired product distills at 190-200" C. under 1 mm. pressure. The product, 4,4'-dimethoxy-a-ethyl chalcone, where R is the ethyl group (see Step I below) is a bright yellow oil.

10 grams of the 4,4'-dimethoxy-a-ethyl chalcone from the previous procedure is dissolved in 50 cc. of absolute alcohol, 1 gram of a copperchromium oxide catalyst added, and the mixture submitted to high-pressure hydrogenation at 220 and about 150 atmospheres pressure for Shows in an appropriate machine. Aiter coolinl. the hydrogen pressure is released and the solution removed irom the hydrogenation apparatus and illtered tree of the catalyst. On cooling the filtrate, the product, 1,3-di-(p-methoxyphenyl) -2-ethylpropane, where R is the ethyl roup crystallizes as a white solid which may be filtered oil. It has a melting point 01 43. By this method of hydrogens-tion, both the double bond and the ketone group in the starting material are completely re duced '(see Step 11 below). Ii desired, the prodnot from this hydrogenation may be converted into the corresponding phenolic product as in Example I (see Step III below).

Bur I (moo-cw cmoOc-cmm 1!! A pmoOc-own-O-oom ii. Sm -II omoO-i-ron-Om on. was. c

cmoocnrimcnoo cm I care in Step I to select the proper ketone for combination with the aldehyde.

Example its-Production of Type 'III Specific members of the series illustrated by Type III may be prepared in the following manner: The first step in this preparation is identical with the first step' oi Example 2, for the production of 4,4'-dimethoxy-a-ethyl chalcone (see Step I below).

The chalcone as produced above may be reacted with ethyl magnesium bromide in the following manner: A solution 01' 8 grams of the chalcone in 150 cc. of dry ether is added slowly to a vigorously stirred solution of 13.2 grams of ethyl magnesium bromide in 85 cc. dry ether maintained at a temperature of --10 C. Stirring is continued for three hours at room temperature after which the reaction m'ixlmre is poured into 500 cc. of ice water containing '15 cc. of hydrochloric acid. The ether layer is separated, the aqueous solution extracted once with ether and the ether solutions combined. After evaporation of the ether the residue is distilled under high vacuum. The distillate, 1,3-dl-(pmethoxyphenyD-2-ethyl pentanone-l, where R and R1 are ethyl radicals, is a pale yellow oil (see Step II below). This may be further converted as follows:

10 grams 0! the ethylpentanone as prepared in Step II is dissolved in 50 cc. of absolute alcohol, 1 gram of a copper-chromium oxide catalyst added, and the mixture submitted to high pres.- sure hydrogenation at 220 and about 150, at-

mospheres pressure'tor 5 hours in an appropriate ll mstepsm and IV.

machine. After cooling, the hydrogen pressure is released and the solution removed from the 'hydrogenstionapparatus and altered tree oi the catalyst. The alcohol is removed from the illtrate by evaporation and the product (see Step III below), which is l,3-di(p-methoxyphenyl)- z-ethylpentaue, where R and R1 are ethyl radicals, is distilled at -175 C. under 1 mm. pressure. It is a colorless liquid and may be converted into the corresponding phenolic compoun asin Example! (seeStepIVbelow).

PIODUC'I'IOI or Trn III Step I Step II cmoO-i-rcn-O-ocm RiMgBr CmoOi-iH-ifi-Oo on,

ste in t omoOi-in-iaOocm (reduction) 1 omoOcn in-inO-oom R and R1 may be the same or diilerent alkyl radicals. In general any desired compound corresponding to this type formula may be made by using, in Step I, a ketone having the proper alhl radical and, in Step II a magnesium bromide compound having the proper alkyl radical.

Since the carbon atoms to which the alkyl radicals are attachedare asymmetric in this type of compound, the product produced may be a single racemate or a mixture 01' racemates and the individual racemates 01 such mixture may be separated by any suitable method, a preferred method being disclosed in our copending application Serial No. 538,090 flied May 30, 1944. As also stated in that application, an individual racemic mixture of two stereoisomeric forms or this type of compound may have greater estrogenic potency than that of the conglomerate oi stereoisomers. For example, when R. and R1 are ethyl groups,"the intermediate compound produced as shown in Step II is in the form of two racemic mixtures, one liquid and one solid. These mixtures .may be separated by dissolving them in a solvent such as alcohol and crystallizing the solid. racemic mixture from the solution. By evaporating the solvent from the mother liquid, the liquid racemic mixture may be recovered. Either the solid or the liquid racemic mixture may then be subjected to the reactions shown Example 4.Production of Type IV Specific members of the series illustrated by Type IV may be prepared in the following manner: p,p'-dlmethoxy-chalcone is prepared in exactly th manner described in Example I (see Step Ibelow). This is thei. treated as follows:

Following exactly the same directions as described in Example 1, Step II, the p,p'-dlmethoiw chalcone is converted into LS-dKp-methoxyphenyl) -pentanone-1 (see Step II below). This is then treated according to Step III as follows:

A solution of grams of the pentanone in 130 cc. dry ether may be added slowly with vigorous stirring to a solution of 8.9 grams of ethyl magnesium bromide in 60 cc. dry ether. After the addition or the pentanone has been completed, the mixture is warmed on a steam bath for five hours, after which the mixture is poured into a mixture of 700 cc. ice water and 100 cc. hydrochloric acid. The ether layer is separated, and the ether removed. The residue is subjected to vacuum distillation, and is obtained as a colorless oil which has a boiling point of 183-184 C. at 2 mm. and which solidifies to a solid compound melting after. purification at 87 C. During this distillation water is evolved and the resulting product is 3,6-di(p-methoxyphenyl) -heptene-2,

where R and R1 are ethyl radicals, (see Step III below). This is then treated according to Step IV as follows:

4 grams of the 3,5-di(p-methoxyphenyl) heptene-2 is dissolved in 70 cc. of glacial acetic acid and 0.04 grams of platinum oxide catalyst added. The container is then attached to a machine for catalytic hydrogenation, the hydrogen applied, and the mixture shaken until one molecular equivalent of hydrogen has been consumed.

The mixture is then decanted from the catalyst, diluted with water, neutralized with NaOH, and the product extracted with ether. The ether is evaporated at room temperature, and the residue distilled under reduced pressure. The product thus obtained (see Step IV below) is a colorless oily liquid and is 3,5-di(p-methoxy-phenvl)- heptane, where R and R1 are ethyl radicals, and may be converted into the corresponding phenolic compound as in Example 1 (see Step V below).

Pxonucrron or TYPI: 4

Step I Step II Step III omo-OE-cm-Zn-Oocm 21mm CHaOO-E-CHrEH-O-OCH;

Step II CHsO- -E-CHs-EH- -0 GB: hydrogenation cnwO-Zn-cm-Zn-O-o on.

Step v' omoQem-cnlcnGocm Bl it R and R1 may be the same or diiferent alkyl radicals. In general, any compound conforming to thi type formula may be made by using in Steps II and III magnesium bromide compounds'having alkyl radicals of the constitution desired in the final product.

Since the carbon atom to which the alkyl radicals are attached are asymmetric in this type of compound, the product may be a single racemate or a mixture of racemates and the individual racemates of such mixture may be separated.

Example 5.Pr0duction of Type V Specific members of the series illustrated by Type V may be prepared in the following manner: The first two steps in this process are identical in all respects with the first two steps in Example 3 for the production or l,3-di(p-methoxyphenyl) -2-ethyl pentanone-l (see Steps I and 11 below). This may be converted into other products as follows:

8.0 grams of the ethyl pentanone obtained in Step II are dissolved in dry ether and added slowly to a solution of ethyl magnesium bromide prepared from 5.0 grams oi ethyl bromide and 1.2 grams of magnesium tumings in anhydrous ether. When the addition or the pentanone has been completed. the reaction mixture is warmed on a steam bath for five hours after which it is poured into a mixture of 700 cc. ice water and cc. hydrochloric acid. The ether layer is separated, dried, and the ether removed. The residue is subjected to vacuum distillation, and is obtained as a colorless oil. During this distillation, water is evolved, and the resulting unsaturated product is 4-ethyl-3,5-di(p-methoxyphenyl)-heptene-2, where R1 and R: are ethyl group (see Step III below).

The 4 ethyl 3,5 dl(p-methoxyphenyl) -heptene-2 from the previous step is dissolved in 70 cc. glacial acetic acid and subjected to catalytic hydrogenation as described in Example 4, Step IV, until no more hydrogen is consumed. The mixture is then decanted from the catalyst, diluted with water, neutralized with sodium hydroxide, and the product extracted with ether. The ether is evaporated at room temperature, and the remaining liquid distilled under reduced pressure. The product thus obtained (see Step IV below) is a colorless oily liquid and i 4-ethyl- 3,5-di(p-methoxyphenyl)-heptane, where R, R1 and R: are ethyl radicals, and may be converted into the corresponding phenolic compound, as in Example 1.

2.00am Paowmcr'rmli Inthepreeentepeclucationandclaimsthe smr m Q OHQWK.

omoOommmoGc-cmm 5 i a Y a a Step, in

cmo-Oi-inchOoonm RMIBI v j R:

cmo-Oi-fln-i Oocn cmo-Oins-in-Zn-Oo cm noO-in iH-EnOon R, R1 and Re maybe the same or diil'erent m1] radicals. In-general. any compound corresponding tothis type'iormula may be made by employing in Step 1- av ketone having the desired alkyl radical, and in Steps II and III by. employing magnesium compounds having thedesired alkyl radicals. Mixtures of the racemates produced I may be separated into the individual racemates.

e. g. by the method-disclosed in the copending application hereinabove referred to.

In the above reactions the terminal group was methoxy, but it may. be any. alkoxy group, and may be obtained in the final product, by the use of appropriate starting compounds in Step I or by converting the OH groups in the final phenolic compound. When a terminal acyl group isvdesired in the final compound it may be conveniently obtained by converting the OH groups in the final phenolic compound into R000 groups. The terminal groups may be in the ortho, meta or para position, symmetrical or not. -'1o produce terminal groups which are esters of inorsanic acids, the phenolic terminal group is condensed with an appropriate inorganic acid; such as phosphoric, and sulionic acid. Ethyl magnesium bromide was used. in each of the foregoing-w actions and excellent results are obtained by its use, but it can be replaced by any alkyl magnesium halide except the fluoride.

. .The compounds or this invention are pretersbly administered by intramuscular injection eimifleethenucleus. I

The dihydroxy compounds of this invention havethegeneral formulait a a on D 0 i i i Where R is selected from the group consistinz of hydrogen and alkyl radicals, at least one of saidRsubetitutesbeinganalkylradical. The hydroxy substituente may be in the ortho, meta.

or para position. and symmetrical or unsym By substituting meta-methoxyacetophenone for the para compound in Example 1 on paged andexactlyasprescribedinthatexample. there is obtained no n H] can th gh I By substituting ortho-methoxyacetophenone for the para compound in Example 1 and proceeding otherwise exactly as in that example, there is obtained H0 H: n elm i G 0 By substituting ortho-methoxybenzaldehyde tor the para compound in Example 1 and n- 'propyl' magnesium bromide for ethyl magnesium bromide and proceeding otherwise exactly as in that example, there is obtained n n on on I I 7 MOL t Example 3 shows the production of By substituting 'ortho-methoxybehzaldehyde for the para compound in Example 3 and otherwise proceeding exactly as in that example, there is obtained 7 Calls Q R Example 4 shows the production of H H 11 g Z' cm1coo-O o-C -oooc.n1 H|(!3 011 x cm. can am H 1 a 5 By substituting ortho-methoxybenzaldehyde f f for the para compound and proceeding other- O O wise exactly as shown in that example, there is 0. obtained on 1-1 11 II I E I o.rH..ooo-C -c--+-c 000cm (E) H0O c lHl C2115 (3H1 i ii. 1 Example 5 shows the production 01' 15 E E 1 0.11,.000 000cm cm. can can 0 0 no t +-o-Oon III H {I 11 Int: III By substituting ortho-methoxybenzaldehyde for cdhcoo-Qd-c-c-Q-moocun the para compound and methyl magnesium iodide for ethyl magnesium bromide in Step III of that example and otherwise proceeding exactly as shown in Example 5, there is obtained H Hall I This compound possesses estrogenic activity of the same order of magnitude as the corresponding di-para compound.

In so far as the esters are concerned, they may be obtained, in general, by esterifying one or both of the hydroxy groups in accordance with the esterification technique known to organic chemists, and the invention includes generically the ester derivatives of organic acids. Support for this generic statement will be found in the following illustrative species of ester derivatives:

CH CaHt H H c,H.eoo-O +-t 00000.11.

H: H H: CHICOOOPJ-fiE- -QOCOCHI Esters such as those listed above may be prepared by any of the usual esteriflcation methods. The following procedures are representative examples:

One gram of 1,3-di-(p-hydroxy phenyl)-nhexane is dissolved in 5 cc. of benzoyl chloride and to this solution one drop of concentrated sulfuric acid is added. After standing several hours, the mixture is diluted with Water, made alkaline with potassium hydroxide and extracted with ether. The ether solution is separated and after boiling oi! the ether, the residue is crystallized from alcohol. The dibenzoate is a white crystalline solid melting at C.

One gram of 1,3-di-(p-hydroxy phenyl)-2- ethyl pentane is refluxed for four hours with a mixture of 10 cc. of acetic anhydride and 0.5 ram of fused sodium acetate. After dilution with water, the mixture is made alkaline with potassium hydroxide, and extracted with ether. The ether solution is separated, the ether boiled off and the residue distilled under high vacuum, It is a thick, pale yellow oil.

Three grams of paimityl chloride is added dropwise to a solution of 1 gram of 2,4-di-(p-livdroxy phenyD-3-ethyl-hexane in 10 cc. of pyridine. After standing overnight, the reaction mixture is worked up in the usual manner. The dipalmitate, after crystallization from alcohol, is a white crystalline solid melting at 36-38 C. All of the above esters have estrogenic properties.

As for the ether derivatives, the invention includes generically the monoor di-alkyl ether derivatives 01' the hydroxy compounds, Support for this generic statement is found in the foregoing description, in which it is pointed out that instead of employing in Step I methoxy compounds, alkoxy compounds in general ma be substituted, and that these alkoxy groups may be in the ortho, meta, or para position in relation to the other substituents, thus producing as a final step prior to hydrolysis, alkoxy or ether derivatives in general, symmetrical and unsymmetrical. Instead of producing alkoxy or other derivatives by starting with appropriate alkoxy compounds, it is entirely possible to convert the dual dihydroxy derivatives into their corresponding ethers by known etheriflcation methods.

n, n, n cmmO -E-im-O-oclm cw lc CIHI 1H:

CsHuO 0 5111:

H; H H, O 0

As previously pointed out, there is an R. substituent at one or more of the three carbon atoms of the propane bridge connecting the benzene radicals, and that R substituent is an alkyl radical in general, and those R substituents are the same or diflerent alkyl radicals in general. Further support for these generic statements will be found in the following specific examples:

Q -ijfG H0 i i-ELK ('31! Ch; g5)

no v on O yc no Li OH m am. $59

It is preferred that the number of carbon atoms in each or the alkyl substituents on the propane bridge be not greater than live, although the activity of such compounds having longer substituent chains has been proved by many experiments, numerous examples of which are found herein.

We claim:

1. Alpha, gamma-iii (hydroxyphenyl) propanes in which at least one of the propane carbon atoms is substituted by an alkyl group.

2. Alpha, gamma-d1 (hydroxyphenyl) propanes in which at least one of the propane carbon atoms is substituted by an alkyl group of less than six carbon atoms.

3. 3-Ethyl-2,4-di (p-hydroxyphenyl) hexane.

4. A racemic mixture of two stereoisomeric forms oi 3-ethyl-2,4-di (p-hydroxyphenyl) hexane havin: estroaenlc potency higher than the conglomerate o1 stereoisomers oi! this compound. 

